GABRG2 Deletion Linked to Genetic Epilepsy with Febrile Seizures Plus Affects the Expression of GABA A Receptor Subunits and Other Genes at Different Temperatures
GABRG2 Deletion Linked to Genetic Epilepsy with Febrile Seizures Plus Affects the Expression of GABA A Receptor Subunits and Other Genes at Different Temperatures
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DOI：
10.1016/j.neuroscience.2020.04.049
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发表时间：
2020-07-01
期刊：
影响因子：
3.3
通讯作者：
Sun, Tao
中科院分区：
文献类型：
--
作者：
Li, Xinxiao;Guo, Shengnan;Sun, Tao
γ-氨基丁酸A受体（GABA（A））亚单位和钠通道基因，尤其是GABRG 2和SCN 1A的突变与热性惊厥（FS）和遗传性癫痫伴热性惊厥加重（GEFS+）有关。GEFS+是一个常染色体显性遗传的儿童癫痫综合征家族。其最常见的表型是热性惊厥，通常伴有辅助性无热性全身强直阵挛性惊厥、热性惊厥+（FS+）、严重癫痫性脑病以及其他类型的全身性或局部相关性惊厥。然而，热性惊厥的发病机制在很大程度上仍不清楚。在此，我们通过在HT-22小鼠海马神经元细胞系中应用CRISPR/Cas9介导的基因组缺失来产生GABRG 2基因敲除细胞系（HT 22（GABRG 2）（KO）），以在体外探索GABRG 2的功能。通过mRNA-seq，我们发现当GABRG 2被敲除时，几个癫痫相关基因的表达谱发生了显着变化，其中一些也显示出温度诱导的变化。京都百科全书基因和基因组（KEGG）分析揭示MAPK和PI 3 K-Akt信号通路的显著改变。我们还观察到GABRG 2敲除后基质金属蛋白酶（MMPs）家族的上调。此外，随着温度的升高，GABRA 1和CACNA 1A（而不是其他）的表达显著降低是一个新的发现。总之，GABA（A）受体突变可导致受体数量减少，这可能导致GABA能通路信号传导受损。该数据首次揭示了GABRG 2突变会影响其他基因的功能，基于这一发现，我们希望这项工作也能为GEFS+中GABRG 2的研究提供新的方向。本研究为癫痫的严重程度提供了分子基础，并为临床治疗癫痫提供了以GABA（A）受体为靶点的药物，为癫痫的基因诊断和靶向治疗提供了新的策略。
Mutations in gamma-aminobutyric acid A receptor (GABA(A)) subunits and sodium channel genes, especially GABRG2 and SCN1A, have been reported to be associated with febrile seizures (FS) and genetic epilepsy with febrile seizures plus (GEFS+). GEFS+ is a well-known family of epileptic syndrome with autosomal dominant inheritance in children. Its most common phenotypes are febrile seizures often with accessory afebrile generalized tonic-clonic seizures, febrile seizures plus (FS+), severe epileptic encephalopathy, as well as other types of generalized or localization-related seizures. However, the pathogenesis of febrile seizures remains largely unknown. Here, we generated a GABRG2 gene knockout cell line (HT22(GABRG2) (KO)) by applying the CRISPR/Cas9-mediated genomic deletion in HT-22 mouse hippocampal neuronal cell line to explore the function of GABRG2 in vitro. With mRNA-seq, we found significant changes in the expression profiles of several epilepsy-related genes when GABRG2 was knockout, some of them showing temperature-induced changes as well. Kyoto Encyclopedia Gene and Genomic (KEGG) analysis revealed a significant alteration in the MAPK and PI3K-Akt signaling pathways. We also observed an up-regulation of the matrix metalloproteinases (MMPs) family after GABRG2 knockout. Furthermore, the significant decrease in expression of GABRA1 and CACNA1A (but not others) with an increase in temperature is a novel finding. In summary, mutations in the GABA(A) receptor can lead to a decrease in numbers of receptors, which may cause the impairment of GABAergic pathway signaling. This data has been the first time to reveal that GABRG2 mutations would affect the function of other genes, and based on this finding we hope this work would also provide a new direction for the research of GABRG2 in GEFS+. It also may provide a molecular basis for the severity of epilepsy, and guide the clinical medication for the treatment of the epilepsy focused on the function on GABA(A) receptors, which, might be a new strategy for genetic diagnosis and targeted treatment of epilepsy.